Cornell iGEM understands the inherent risks of working in a lab facility and aims to take all necessary precautions to ensure no personal or environmental harm occurs. To this end, we have implemented the following safety procedures below. Our completed safety form can be found here.
Specific Safety Concerns
Laboratory Safety: Our project involves regular use of ethidium bromide, a DNA-intercalating agent known to cause cancer, as well as the use of powerful UV light, for visualization of gel electrophoresis. We must prepare culture media with antibiotics, which could be harmful to humans in large doses. We also work with ethanol lamps to maintain a sterile environment, which do involve having an open flame on the benchtop.
Environmental Safety: If any biological materials escape from the lab there is a risk of transfer of antibiotic resistance from our engineered strains into other organisms. Furthermore, dissemination of the peptides part of our product could potentially affect the microbiological ecosystem present at the release point, but we plan to secure the release of the peptide using a commonly found syringe specifically for injection of medication to the cow’s teats.
Pathogens in Lab: We are mainly working with S. aureus, S. epidermidis, E. coli, C. freundii, K. pneumoniae in the lab space. We have chosen these variety of pathogens in order to have a variety of Gram Negative and Gram Positive bacteria to test the bacteriocins effectiveness against. S. aureus and S. epidermidis are both naturally found on human skin [1,2]. E. coli is found in the human gut flora, but also has been widely used by iGEM teams for cloning purposes. C. freundii is naturally found in the soil and but can also be pathogenic when found in humans. C. freundii, E. coli, and S. epidermidis are all categorized as Biosafety Level 1. However, K. pneumoniae and S. aureus are Biosafety Level 2, and we have had to take significant precautions working with this organism, including working in a biocabinet cabinet and carrying out extra sterilizing procedures to appropriately dispose of biohazards.
Bacteriocins in Lab: We have been working with Bactofencin A, Enterocin E760, Colicin M, and Lysostaphin. These various antimicrobial peptides are bacteriolytic and thus are toxic to specific bacterial cells. Bacteriocins have been known to be harmless to humans as many bacteriocins are naturally found in the human gut and in our everyday environment. Thus, the peptides we are working with are relatively risk-free and do not require extra precaution outside of basic lab safety.
Wet Lab: All lab members wear nitrile gloves, closed-toe shoes, and use eye protection when working with volatile chemicals or UV light. Gloves are replaced and hands are washed immediately after using ethidium bromide or any of the metal solutions. Members work in small groups to ensure if any harm comes to one, others are there to assist. When working with a new reagent or piece of equipment, a faculty lab manager or experienced member is always present to assist.
There are taped off, designated areas for working with ethidium bromide. These areas are cleaned before and after work and are the only areas the solution may touch. All toxic waste is placed in a specialized receptacle and is picked up and disposed of by Cornell Environmental Health and Safety.
All disposables that come in contact with biologics are disposed of in biohazard waste. The lab space also contains sharps containers for disposal of all sharps that contact biological material. All biohazard waste is autoclaved and transported to the building's centralized waste facility where it is disposed of as regulated biological waste.
We maintain 2 copies of MSDS's for every chemical we use in the lab: one for our own records and one for the lab manager and users of the lab space who are not part of our team. The lab is equipped with flame-retardant benches, spill kits, safety showers, eye-washes, and fire extinguishers.
Dry Lab: We use the Emerson Machine Shop for fabrication; each of the dry lab subteam members has attended the prescribed training session for use of the shop and has learned to use each of the tools safely. Each member of the dry lab subteam was trained in the safe usage of the milling machine and the metal lathe.
All machine shop work is conducted under the supervision of the Emerson machine shop staff. Safety goggles were worn at all times. Masks and gloves are worn as appropriate. Closed-toe shoes and long pants were also worn when working in the machine shop. While working in the machine shop we maintained a clean work environment so we could maintain visibility at all times. When lifting heavy objects, proper lifting technique was used, and an appropriate number of individuals were used for lifting said objects.
Training and Enforcement
Training: All team members who work in the wet lab must complete Cornell EH&S general lab safety and chemical waste disposal courses prior to the onset of work. These courses set specific guidelines and are the standard requirement for work in a biosafety-level 1 lab at Cornell. Additionally, all team members must complete a lab orientation session with the manager of the BME instructional lab, Dr. Shivaun Archer. During these sessions, Dr. Archer familiarizes new members with the safety equipment and procedures specific to the labspace in which we work.
Prior to the onset of work for the year, all new members are required to go through a safety training program. During this program, safety officers reinforce safety procedures learned during the EH&S courses, discuss safety protocol pertaining to specific chemicals with which we work, and ensure all lab members fully understand all safety procedures.
Safety Officers: The safety officers were team members chosen to directly supervise the activities of the other team members. One team member each was chosen for the wet and dry lab subteams to ensure that all team members were working safely, whether with bacterial cultures or power tools. These team members also act as liaisons to the wet lab and machine shop managers and, when necessary, the Institutional Biosafety Committee to ensure proper equipment usage.
These team members are responsible for discussing the proposed work plan for the project with the wet lab and machine shop managers before starting work to ensure that it is safe to continue. In the case of the wet lab in particular, this involves going through a detailed list of protocols, including all organisms, chemicals, and genetic constructs being worked with, to ensure conformity with the Environmental Health & Safety guidelines. They must go through the same safety training as all other team members, but are required to redo the training each time we recruit new members in order to keep up-to-date with safety considerations. In addition, they maintain contact with the supervisors of the workspaces, usually in the form of a weekly check-in, to discuss any safety concerns that have arisen and ensure that equipment continues to be used properly.
Enforcement: Team members who violate safety rules are required to work under the supervision of the safety officers for the remainder of the week, or until the safety officer believes the member is capable of performing the task unsupervised. For multiple infractions or complete disregard to safety protocols, a member may be restricted from laboratory work until he/she undergoes EHS chemical safety online training again, and demonstrates proper performance to a team leader of failed technique(s) in a controlled setting.